In vivo fx pro, by Carestream - Product details

1

Topical Application of Rho-SPIONs and Doxorubicin

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Mice were anesthetised by intraperitoneal injection of ketamine (75 mg/kg) and medetomidine (1 mg/kg), placed on a warming pad and the back skin of the mice was cleaned with a 70% isopropyl alcohol swab (Livingstone International, Australia). Rho-SPIONs (20 µl of 2 mg/mL in PBS) were applied to the skin and left for 1 hour, after which excess liquid was wiped off with a wet cotton bud. PBS only (20 µL) was used for the control mouse. For chemotherapeutic drug delivery, a 30 µL mixture in PBS containing either both 10%NH 2 -SPIONs (1.3 mg/mL as Fe) and DOX (0.17 mM), 10%NH 2 -SPIONs only (1.3 mg/mL as Fe), or DOX only (0.17 mM), was applied to the mouse skin and then left for 1-2 hours, at which time unabsorbed material was removed by wiping. Anaesthesia was reversed with atipamezole (0.2 mg/kg).
The mice were imaged under anesthesia on a Carestream In-Vivo FX PRO (Carestream Health, Woodbridge, USA) using 550 nm excitation and 600 nm emission wavelength lters before application and 1, 2, 24 and 48 hours after application. They were then euthanized, and treated skins were xed in 4% paraformaldehyde (PFA) overnight at 4 o C for uorescent imaging.

Raviraj V., Pham B.T.T., Pham N.T., Kim B.J., Kok L.F., Painter N., Delic N.C., Jones S., Hawkett B.S., & Lyons J.G. (2020). Non-Invasive Transdermal Delivery of Chemotherapeutic Molecules In Vivo Using Superparamagnetic Iron Oxide Nanoparticles.

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Topical Application of Rho-SPIONs and Doxorubicin

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Mice were anesthetised by intraperitoneal injection of ketamine (75 mg/kg) and medetomidine (1 mg/kg), placed on a warming pad and the back skin of the mice was cleaned with a 70% isopropyl alcohol swab (Livingstone International, Australia). Rho-SPIONs (20 µl of 2 mg/mL in PBS) were applied to the skin and left for 1 hour, after which excess liquid was wiped off with a wet cotton bud. PBS only (20 µL) was used for the control mouse. For chemotherapeutic drug delivery, a 30 µL mixture in PBS containing either both 10%NH 2 -SPIONs (1.3 mg/mL as Fe) and DOX (0.17 mM), 10%NH 2 -SPIONs only (1.3 mg/mL as Fe), or DOX only (0.17 mM), was applied to the mouse skin and then left for 1-2 hours, at which time unabsorbed material was removed by wiping. Anaesthesia was reversed with atipamezole (0.2 mg/kg).
The mice were imaged under anesthesia on a Carestream In-Vivo FX PRO (Carestream Health, Woodbridge, USA) using 550 nm excitation and 600 nm emission wavelength lters before application and 1, 2, 24 and 48 hours after application. They were then euthanized, and treated skins were xed in 4% paraformaldehyde (PFA) overnight at 4 o C for uorescent imaging.

Raviraj V., Pham B.T.T., Kim B.J., Pham N.T., Kok L.F., Painter N., Delic N.C., Jones S.K., Hawkett B.S., & Lyons J.G. (2021). Non-Invasive Transdermal Delivery of Chemotherapeutic Molecules In Vivo Using Superparamagnetic Iron Oxide Nanoparticles.

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3

Topical Nanoparticle-Mediated Delivery

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Mice were anesthetized by intraperitoneal injection of ketamine (75 mg/kg) and medetomidine (1 mg/kg), placed on a warming pad and the back skin of the mice was cleaned with a 70% isopropyl alcohol swab (Livingstone International, Australia). Rho-SPIONs (20 µl of 2 mg/mL in PBS) were applied to the skin and left for 1 h, after which excess liquid was wiped off with a wet cotton bud. PBS only (20 µL) was used for the control mouse. For chemotherapeutic drug delivery, a 30 µL mixture in PBS containing either both 10% NH 2 -SPIONs (1.3 mg/mL as Fe) and DOX (0.17 mM), 10% NH 2 -SPIONs only (1.3 mg/mL as Fe), or DOX only (0.17 mM), was applied to the mouse skin and then left for 1-2 h, at which time unabsorbed material was removed by wiping. Anesthesia was reversed with atipamezole (0.2 mg/kg).
The mice were imaged under anesthesia on a Carestream In Vivo FX PRO (Carestream Health, Woodbridge, USA) using 550 nm excitation and 600 nm emission wavelength filters before application and 1, 2, 24 and 48 h after application. They were then euthanized, and treated skins were fixed in 4% paraformaldehyde (PFA) overnight at 4 °C for fluorescent imaging.

Raviraj V., Pham B.T.T., Kim B.J., Pham N.T., Kok L.F., Painter N., Delic N.C., Jones S., Hawkett B.S., & Lyons J.G. (2021). Non-invasive transdermal delivery of chemotherapeutic molecules in vivo using superparamagnetic iron oxide nanoparticles. Cancer Nanotechnology, 12(1).

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4

Tissue Distribution of DiR-Labeled DPT-PM

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For tissue distribution study, Hela229 tumor cell line (5 × 10⁵ cell/200 μL) was injected into the right-limb flank of the nude mice (weight : 20±2 g, n = 12). After 2 weeks of tumor growth, all mice were divided into two groups (n = 6). One group was injected with 200 μL of DiR-labeled DPT-PM via tail vein and another group was injected with free DiR at a fixed DiR dose of 1.25 mg/kg in both groups. After injection, both groups were imaged under in vivo imaging system (In-Vivo FX PRO, Carestream, Canada) at the time interval of 1, 2, 4, 6 and 8 h. After 8 h of imaging, one mouse from each group was sacrificed to harvest the tumor and major organs (heart, liver, spleen, lung and kidney) and was imaged under IVIS and the obtained fluorescence intensities were calculated.

Zu C., Yu Y., Yu C., Li Y., Sun R., Chaurasiya B., Tang B., Chen D., Tu J, & Shen Y. (2020). Highly loaded deoxypodophyllotoxin nano-formulation delivered by methoxy polyethylene glycol-block-poly (D,L-lactide) micelles for efficient cancer therapy. Drug Delivery, 27(1), 248-257.

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5

Biodistribution of DiR-Labeled GlcA-NPplex in MCT-PH Rats

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Free DiR or DiR-GlcA-NPplex was injected into MCT-PH rats via the tail vein. The rats were sacrificed 24 h after the administration by neck dissection. Then the major tissues, including the heart, liver, spleen, lung, and kidney, were dissected for ex vivo imaging (KODAK, In-Vivo FX PRO, Carestream, Canada). The excitation and emission wavelengths of the DiR channel are 748 and 780 nm, respectively.
For the biodistribution study, the MCT-PH rats (3 rats/group) were intravenously injected with DiR-labeled GlcA-NPplex via the tail vein at a DiR dose of 0.5 mg/kg, according to the rats' body weight. The co-localization of the GlcA-NPplex with PA α-SMA (α-smooth muscle actin) in the models was studied using Leica Corp. IX 7 fluorescence microscopy (Leica Microsystems Inc. IX7, Wetzlar, Germany). The rats received intravenous administration of the free IR783-labeled preparations at 0.5 mg/kg IR783. Four hours after the treatment, the rats’ lungs were isolated and embedded with OCT for further fluorescent imaging. Briefly, the lung sections were cut into 2 μm and then blocked for 15 min with 5% FBS at room temperature. After the excess-serum removal, the sections were stained for 12 h with a mouse anti-rat α-SMA monoclonal antibody at 5 μg/mL and incubated with FITC (diluted 1:200 with FITC buffer) for 1 h.

Li B., Teng C., Yu H., Jiang X., Xing X., Jiang Q., Lin C., Zhao Z., Zhang R, & He W. (2022). Alleviating experimental pulmonary hypertension via co-delivering FoxO1 stimulus and apoptosis activator to hyperproliferating pulmonary arteries. Acta Pharmaceutica Sinica. B, 13(6), 2369-2382.

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6

In Vivo Bioluminescence Imaging

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After transfection with plasmid or Doggybone encoding luciferase, expression was visualized using an In vivo FX pro (Carestream, USA) following intraperitoneal injection of Rediject D-Luciferin (Caliper) in accordance with manufacturer’s specification. Light emission was measured for 4 minutes without binning and an X-ray was taken for 30 seconds, the two images were overlaid using Carestream MI SE software. Relative luminescence was quantified by using the software’s region of interest (ROI) analysis function, with background levels set using control, untransfected animals.

Walters A.A., Kinnear E., Shattock R.J., McDonald J.U., Caproni L.J., Porter N, & Tregoning J.S. (2014). Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines. Gene therapy, 21(7), 645-652.

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7

Biodistribution of Tumor-Targeted DiR Agents

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The BALB/C mice were inoculated subcutaneously under the axilla with 1 × 10⁶ suspended 4T1 cells. The experiment began at 2 weeks after tumor cells were injected. 200 μL of PTX@TF/DiR-LTSLs or DiR-LTSLs or free DiR was injected into the mice at a fixed DiR dose of 0.5 mg/kg via the tail vein. At 0.5, 1, 2, 4, 8 and 24 h after injection, fluorescence images were obtained by an in vivo imaging system (IN-VIVO FX Pro, Carestream, Canada) after mice were anesthetized. Finally, the mice were put down for their major organs and tumors. Fluorescence intensity in various organs was systematically calculated by an in vivo imaging system.

Zhang K., Li J., Xin X., Du X., Zhao D., Qin C., Han X., Huo M., Yang L, & Yin L. (2021). Dual Targeting of Cancer Cells and MMPs with Self-Assembly Hybrid Nanoparticles for Combination Therapy in Combating Cancer. Pharmaceutics, 13(12), 1990.

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8

In Vivo Fluorescence Imaging of Grafted Cells

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FLI was performed on a small animal in vivo FLI system (In-Vivo FxPro; Carestream, MI, USA) immediately after MRI. White light imaging, FLI with 487-nm excitation wavelength and 509-nm emission wavelength and digital X-ray imaging were obtained to detect the grafted cells. The fluorescence intensity of the in vivo imaging tests was quantified using the Carestream MI software by an author (M.C.), who was blinded to the experimental groups.

Zhang F., Duan X., Lu L., Zhang X., Chen M., Mao J., Cao M, & Shen J. (2017). In Vivo Long-Term Tracking of Neural Stem Cells Transplanted into an Acute Ischemic Stroke model with Reporter Gene-Based Bimodal MR and Optical Imaging. Cell Transplantation, 26(10), 1648-1662.

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9

In Vivo Bioluminescence Imaging

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D-Luciferin (Biosynth, Switzerland) was administered i.p. at a dose of 150 mg/kg 10 minutes before imaging. Images were acquired using an In-Vivo FX Pro molecular imaging system (Carestream Health Inc., NY, USA). Analysis was performed with the Carestream molecular imaging software v5.0.6.20.

Ruiz de Garibay G., García de Jalón E., Stigen E., Lund K.B., Popa M., Davidson B., Safont M.M., Rygh C.B., Espedal H., Barrett T.M., Haug B.E, & McCormack E. (2021). Repurposing 18F-FMISO as a PET tracer for translational imaging of nitroreductase-based gene directed enzyme prodrug therapy. Theranostics, 11(12), 6044-6057.

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Bioluminescent Tumor Imaging in Mice

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Mice were imaged once a week, two days after ultrasound treatment, using an In-Vivo FX PRO molecular imaging system (Carestream Health, Inc., Rochester, NY, USA). The mice were anesthetized using isoflurane in O₂ and 150 mg/kg D-luciferin (Biosynth AG, Staad, Switzerland) was injected intraperitoneally (IP) 10 min prior to imaging. The left flank of the mice was imaged to ensure the complete tumor was captured in a single scan. Total bioluminescence values were measured using manual ROIs with background correction in the Carestream MI software (Standard Edition, v.5.0.6.20, Carestream Health, Inc.). A whole-body ROI was used to include metastatic spread.

Kotopoulis S., Popa M., Mayoral Safont M., Murvold E., Haugse R., Langer A., Dimcevski G., Lam C., Bjånes T., Gilja O.H, & Cormack E.M. (2022). SonoVue® vs. Sonazoid™ vs. Optison™: Which Bubble Is Best for Low-Intensity Sonoporation of Pancreatic Ductal Adenocarcinoma?. Pharmaceutics, 14(1), 98.

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In vivo fx pro

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12 protocols using in vivo fx pro

Topical Application of Rho-SPIONs and Doxorubicin

Topical Application of Rho-SPIONs and Doxorubicin

Topical Nanoparticle-Mediated Delivery

Tissue Distribution of DiR-Labeled DPT-PM

Biodistribution of DiR-Labeled GlcA-NPplex in MCT-PH Rats

In Vivo Bioluminescence Imaging

Biodistribution of Tumor-Targeted DiR Agents

In Vivo Fluorescence Imaging of Grafted Cells

In Vivo Bioluminescence Imaging

Bioluminescent Tumor Imaging in Mice

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